This is about on TLC. and I hope it will helpful for you. In this describe about their introduction, principle, application, procedure, methodology, RF value, and their advantage, disadvantage Thank you😊

1. Tarique Husain
B.Pharm

2. • Thin layer chromatography is a type of liquid
chromatography that is used to separate non-
volatile mixtures such as glycerin into its
individual components.
• The process is performed on a sheet of glass,
plastic, or aluminium foil coated with a thin layer
of adsorbent material like silica gel, aluminium
oxide (alumina), or cellulose (known as the
stationary phase).
• In TLC, components of the mixture are
partitioned between an adsorbent (the stationary
phase, usually silica gel) and a solvent ( the
mobile phase, usually hexane : ethyl acetate).

3. • The principle for separation is
adsorption.
• One or more compounds are spotted
on a thin layer of adsorbent coated on
a chromatographic plate(TLC Plate).
• The M/P flows through because of
capillary action.
• The components move according to
their affinities towards the adsorbent.
More affinities towards the S/P –
travels slowly.
Lesser affinities towards the S/P –
travels faster.

4. THE STATIONARY PHASE THAT IS
APPLIED TO THE PLATE IS MADE TO
DRY AND STABILIZE.
• To apply sample spots, thin marks are
made at the bottom of the plate with
the help of a pencil.
• Apply sample solutions to the marked
spots.
• Pour the mobile phase into the TLC
chamber and to maintain equal
humidity, place a moistened filter paper
in the mobile phase.
• Place the plate in the TLC chamber
and close it with a lid. It is kept in such
a way that the sample faces the mobile
phase.
• Wait till the development of spots.
Once the spots are developed, take
out the plates and dry them. The

5. A:- UNDER UV 254nm
B:- UNDER UV 366nm
C:- UNDER SUNLIGHT

6. • TLC is very commonly used technique in
synthetic chemistry for identifying compound,
determining their purity and the progress of a
reaction.
• Purity of sample
• Examination of reaction
• Identification of compounds
• Separation of multicomponent pharmaceutical
formulation (like vitamins, antibiotics, protein,
glycoside.

8. Rf=
Rf=
distance traveled by solute
(a)
distance traveled by solvent
(b)
a
b
4.5
6
=0.75

9. ADVANTAGES
• An easy method of
separation of the
components.
• Its cheaper
chromatographic technique.
• Greater speed of
separation.
DISADVANTAGES
• Accurate quantitative
analysis may not be
performed by TLC .
• Not suitable for volatile
compound.